CN115501156A - 一种具有美白舒缓功效的化妆品原料组合物及其评价体系 - Google Patents
一种具有美白舒缓功效的化妆品原料组合物及其评价体系 Download PDFInfo
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Abstract
本发明公开了一种具有美白舒缓功效的化妆品原料组合物及其评价体系,包括以下成分:光果甘草根提取物、姜黄根提取物、黄芩根提取物、芍药根提取物、三七根提取物,并从基因、细胞、3D皮肤模型、人体四个层面评价美白、舒缓功效。本发明优点是:结合多种天然化妆品原料,美白的同时,舒缓肌肤,防护外界对肌肤的刺激,对皮肤具有良好的美白、舒缓作用;组合的原料比单组分的功效更佳,具有良好的协同增效作用;对组合原料及含该原料的化妆品构建出了一个多维度的评价体系。
Description
技术领域
本发明涉及化妆品配方技术领域、化妆品功效评价技术领域,具体涉及一种具有美白舒缓功效的化妆品原料组合物及其评价体系。
背景技术
古有“肤如凝脂,肌如白雪”,今有“一白遮百丑”,中国女性对于白皙、红润肌肤的向往和追求一直亘古不变。但纵观多年来美白市场的发展,美白产品普遍存在着安全与功效不可兼得的问题,这也使得天然纯净的护肤理念备受关注,对于产品配方中美白功能性原料的合理搭配日渐重视。
皮肤颜色的差异主要是由黑色素细胞的黑色素生成能力决定的。黑色素的数量、类型、分布对皮肤的颜色起着决定性的作用。在黑色素的形成过程中,酪氨酸酶、多巴色素巨变酶、DHICA氧化酶、内皮素等起了巨大的作用。黑色素的生成机理:皮肤内黑素细胞负责生产黑色素。紫外线可以激活黑素细胞,提高酪氨酸酶的活性。在黑色素细胞中,酪氨酸在酪氨酸酶的作用转变成“多巴”,多巴进一步氧化形成“多巴醌”,多巴醌再经过氧化反应形成黑色素,这些黑色素会通过基底细胞的分裂传递到表皮细胞,再通过新陈代谢排出。当新陈代谢减弱时,黑色素就会蓄积起来,形成色斑、雀斑和暗沉。除了紫外线,其他因素如皮肤炎症(炎症因子、炎症介质等)、压力、废气、活性氧等也会促使黑色素细胞活跃,产生过剩的黑色素。因此,在美白的同时往往需要其他功效的辅助,特别是舒缓,可通过缓解外界对皮肤的刺激,从而减少黑色素的生成。
随着2021版《化妆品监督管理条例》(以下简称“新条例”)的颁布,功效宣称再也不是噱头,需要数据和文献资料支撑,并且受到政府和消费者们的监督。在新条例的规定下,对于化妆品功效评价的要求也越来越高。但现有功效评价体系相对单一,缺乏“功效-验证”的整体设计方案。
发明内容
本发明的第一目的在于提供一种天然来源的美白舒缓的化妆品原料组合物,以舒缓助力美白,采用天然来源,顺应市场需求。
采用以下方案实现:一种具有美白舒缓功效的化妆品原料组合物,其特征在于,包括以下成分:光果甘草根提取物、姜黄根提取物、黄芩根提取物、芍药根提取物、三七根提取物;所述光果甘草根提取物占组合物总浓度配比为5%,姜黄根提取物占组合物总浓度配比为5%,黄芩根提取物占组合物总浓度配比为26%,芍药根提取物占组合物总浓度配为11%,三七根提取物占组合物总浓度配比为53%。
进一步的,所述光果甘草根提取物对蘑菇酪氨酸酶活性的IC50为0.1μg/mL,姜黄根提取物对蘑菇酪氨酸酶活性的IC50为2μg/mL,所述两种原料对蘑菇酪氨酸酶活性的抑制作用远大于阳性对照维生素C(对蘑菇酪氨酸酶活性的IC50为50μg/mL)。。
进一步的,黄芩根提取物、芍药根提取物和三七根提取物均可抑制NO的分泌,IC50分别为27μg/mL、41.6μg/mL、190μg/mL,所述三种原料对NO的抑制作用优于阳性对照地塞米松(对NO分泌量的IC50为200μg/mL)。
进一步的,所述光果甘草根和姜黄根提取物由体外蘑菇酪氨酸酶活性抑制试验筛选出;所述黄芩根提取物、芍药根提取物以及三七根提取物由LPS诱导巨噬细胞RAW264.7检测NO含量试验筛选出。
本发明的第二目的是:克服现有技术缺陷,提供一个美白舒缓原料及产品的功效评价体系,从基因、细胞、3D皮肤模型、人体四个层面进行多维度评价,形成“功效-验证”的整体方案,同时,遵循“3R”原则,采用3D皮肤模型来替代传统动物实验。
采用以下方案实现:天然来源的美白舒缓的化妆品原料组合物的评价体系包括:高通量初筛功效模块,用于对所收集的美白、舒缓原料进行初步筛选;基因、细胞功效评价模块,用于美白、舒缓原料组合物复配比例优化及功效评价;3D皮肤模型功效评价模块,用于对含该原料组合物的化妆品成品进行功效评价。
进一步的,所述高通量初筛功效模块,采用体外蘑菇酪氨酸酶活性抑制试验筛选美白原料;采用LPS诱导巨噬细胞RAW264.7检测NO含量试验筛选舒缓原料。
进一步的,基因、细胞功效评价模块,包括对筛选出来的原料通过响应面法进行复配比例优化。
进一步的,所述美白、舒缓原料组合物功效评价具体为:基于α-MSH诱导小鼠黑素瘤细胞B16检测黑色素含量、酪氨酸酶基因表达量试验评价美白功效;基于UVB辐照角质形成细胞HaCat检测ROS含量试验,基于H2O2刺激角质形成细胞HaCat检测SOD、GPx活性和MDA含量试验评价抗氧化功效;基于LPS诱导巨噬细胞RAW264.7检测IL-6、TNF-α、NO含量试验评价舒缓功效。
进一步的,所述3D皮肤模型功效评价模块具体为:基于UVB刺激3D黑素模型检测表观色度、表观亮度试验评价美白功效,基于SLS刺激3D表皮皮肤模型检测IL-1α含量试验评价舒缓功效。
进一步的,还包括人体临床功效评价模块;所述人体临床功效评价模块具体为:基于紫外线诱导人体皮肤黑化模型检测黑素指数、ITA°及视觉肤色评估来进行美白临床功效评价。
本发明的有益效果是:结合多种天然化妆品原料,美白的同时,舒缓肌肤,防护外界对肌肤的刺激,对皮肤具有良好的美白、舒缓作用;组合的原料比单组分的功效更佳,具有良好的协同增效作用;对组合原料及含该原料的化妆品构建出了一个多维度的评价体系。
附图说明
图1黑色素合成测试结果柱状图。
图2酪氨酸酶基因表达测试结果图。
图3 ROS生成量测试结果柱状图。
图4 ROS生成量测试结果图。
图5 SOD活性测试结果柱状图。
图6 GPx活性测试结果柱状图。
图7 MDA含量测试结果柱状图。
图8 IL-6含量测试结果柱状图。
图9 TNF-α含量测试结果柱状图。
图10 NO含量测试结果柱状图。
图11原料组合物与常用美白活性物对黑色素合成抑制率测试结果柱状图。
图12表观色度图片。
图13 L*值检测结果柱状图。
图14 IL-1α含量检测结果柱状图。
图15黑素指数检测结果柱状图。
图16 ITA°检测结果柱状图。
图17肤色检测结果柱状图。
图18美白、舒缓原料组合物及含该原料的化妆品的功效评价流程图。
具体实施方案
以下结合实施例,对本发明进一步详细说明。应当理解,此处所描述的具体实施例仅用以解释本发明,并不用于限定本发明。
本实施例提供一种具有美白舒缓功效的化妆品原料组合物,包括以下成分:光果甘草根提取物、姜黄根提取物、黄芩根提取物、芍药根提取物、三七根提取物;所述光果甘草根提取物占组合物总浓度配比为5%,姜黄根提取物占组合物总浓度配比为5%,黄芩根提取物占组合物总浓度配比为26%,芍药根提取物占组合物总浓度配为11%,三七根提取物占组合物总浓度配比为53%。
此外,本发明对上述组合物制作了一套评价体系,第一部分:美白、舒缓原料的筛选,其中,包括高通量初筛功效模块,具体为:采用体外蘑菇酪氨酸酶活性抑制试验筛选美白原料;采用LPS诱导巨噬细胞RAW264.7检测NO含量试验筛选舒缓原料;最终筛选出来的原料为光果甘草根提取物、姜黄根提取物、黄芩根提取物、芍药根提取物、三七根提取物。
第二部分:美白、舒缓原料复配比例优化,其中,包括对筛选出来的原料通过响应面法进行复配比例优化,得出光果甘草根提取物占组合物总浓度配比为5%,姜黄根提取物占组合物总浓度配比为5%,黄芩根提取物占组合物总浓度配比为26%,芍药根提取物占组合物总浓度配为11%,三七根提取物占组合物总浓度配比为53%。
第三部分:美白、舒缓原料组合物功效评价,其中,包括基因、细胞功效评价模块。具体为:基于α-MSH诱导小鼠黑素瘤细胞B16检测黑色素含量、酪氨酸酶基因表达量试验评价美白功效;基于UVB辐照角质形成细胞HaCat检测ROS含量试验,基于H2O2刺激角质形成细胞HaCat检测SOD、GPx活性和MDA含量试验评价抗氧化功效(提亮肤色);基于LPS诱导巨噬细胞RAW264.7检测IL-6、TNF-α、NO含量试验评价舒缓功效。
第四部分:含美白、舒缓原料组合物的化妆品成品的功效评价,其中,包括3D皮肤模型功效评价模块,具体为:基于UVB刺激3D黑素模型检测表观色度、表观亮度(L*值)试验评价美白功效,基于SLS刺激3D表皮皮肤模型检测IL-1α含量试验评价舒缓功效。
第五部分,含美白、舒缓原料组合物的化妆品成品的功效评价及验证,其中,包括人体临床功效评价模块,具体为:基于紫外线诱导人体皮肤黑化模型检测黑素指数、ITA°及视觉肤色评估来进行美白临床功效评价。
第六部分:美白、舒缓原料组合物及含该原料的化妆品功效评价体系,其中,包括高通量初筛功效模块,基因、细胞功效评价模块,3D皮肤模型功效评价模块,人体临床功效评价模块。
实施例1
高通量初筛功效模块筛选出的美白原料为:光果甘草根提取物、姜黄根提取物,其中,光果甘草根提取物对蘑菇酪氨酸酶活性的IC50为0.1μg/mL,姜黄根提取物对蘑菇酪氨酸酶活性的IC50为2μg/mL。
高通量初筛功效模块筛选出的舒缓原料为:黄芩根提取物、芍药根提取物、三七根提取物,其中,黄芩根提取物对NO分泌量的IC50为27μg/mL,芍药根提取物对NO分泌量的IC50为41.6μg/mL,三七根提取物对NO分泌量的IC50为190μg/mL。
实施例2
响应法优化原料组合物的复配比例,实验结果如表1、2所示。
表1响应法优化美白的复配比例
表2响应法优化舒缓原料的复配比例
以协同增效作用为主要指标,选取表1中的实验编号8,当光果甘草根提取物与姜黄根提取物浓度均为2.5μg/mL时,对B16细胞内黑色素合成抑制率为69.09%,两者协同增效作用达24.92%;选取表2中的实验编号7,黄芩根提取物浓度为12.38μg/mL、芍药根提取物浓度为5.00μg/mL、三七根提取物浓度为25.00μg/mL时,对RAW264.7细胞分泌NO的抑制率为76.87%,三者协同增效作用达65.19%。因此,得出光果甘草根提取物占组合物总浓度配比为5%,姜黄根提取物占组合物总浓度配比为5%,黄芩根提取物占组合物总浓度配比为26%,芍药根提取物占组合物总浓度配为11%,三七根提取物占组合物总浓度配比为53%。
实例3
基因、细胞功效评价模块评估美白、舒缓原料组合物的功效。
美白功效:基于α-MSH诱导小鼠黑素瘤细胞B16通过NaOH裂解法检测原料组合物作用后的黑色素含量,通过qRT-PCR法检测原料组合物作用后酪氨酸酶基因的表达量,实验结果如图1、2所示,原料组合物可显著降低B16细胞内黑色素的合成(P<0.01),可显著下调B16细胞内酪氨酸酶的活性(P<0.05)。
抗氧化功效(提亮肤色):基于UVB辐照角质形成细胞HaCat通过荧光探针检测原料组合物作用后的ROS含量,基于H2O2刺激角质形成细胞HaCat通过WST-8法检测原料组合物作用后的SOD活性、NADPH法检测GPx活性、显色法检测MDA含量,实验结果如图3、4、5、6、7所示,原料组合物可显著抑制ROS的生成(P<0.01),提高SOD、GPx的活性(P<0.01),抑制MDA的生成(P<0.05)。
舒缓功效:基于LPS诱导巨噬细胞RAW264.7通过ELISA法检测原料组合物作用后的IL-6、TNF-α含量、Griess Reagent试剂检测NO含量,实验结果如图8、9、10所示,原料组合物可显著抑制IL-6、TNF-α、NO的分泌(P<0.01)。
功效对比:将原料组合物与常用美白活性物α-熊果苷、传明酸、烟酰胺、AA2G进行功效对比。基于α-MSH诱导小鼠黑素瘤细胞B16通过NaOH裂解法检测活性物作用后的黑色素含量。原料组合物的测试浓度为0.05mg/mL,α-熊果苷、烟酰胺、AA2G的测试浓度均为1mg/mL,传明酸测试浓度为2mg/mL,实验结果如图11所示。原料组合物在较低浓度下对于B16细胞黑色素生成抑制作用优于常用美白活性物α-熊果苷、传明酸、烟酰胺、AA2G。
实例4
3D皮肤模型功效评价模块评估含美白、舒缓原料组合物的化妆品成品的功效,含美白、舒缓原料组合物的化妆品成品剂型为乳液。
美白功效:基于UVB刺激3D黑素模型通过拍照检测乳液作用后的表观色度、色度仪检测表观亮度(L*值),实验结果如11、12所示,含美白、舒缓原料组合物的乳液可显著提高3D皮肤模型的表观色度、表观亮度(L*值)(P<0.01)。
舒缓功效:基于SLS刺激3D表皮皮肤模型通过ELISA法检测乳液作用后的IL-1α含量,实验结果如图13所示,含美白、舒缓原料组合物的乳液可显著抑制IL-1α的分泌(P<0.01)。
实例5
人体临床功效评价模块评估含美白、舒缓原料组合物的化妆品成品的功效,含美白、舒缓原料组合物的化妆品成品剂型为乳液。
美白功效:基于紫外线诱导人体皮肤黑化模型通过皮肤黑素检测仪、皮肤色度仪测量来检测乳液涂抹4周后的黑素指数、ITA°的变化,并由皮肤科医生借助由浅至深肤色的色卡对各测试区肤色进行评估,测试人数30人,实验结果如图15、16、17所示,含美白、舒缓原料组合物的乳液可显著降低人体黑素指数,提高ITA°,改善视觉肤色。
实例6
美白、舒缓原料组合物及含该原料的化妆品功效评价体系,具体流程如图18所示。
Claims (10)
1.一种具有美白舒缓功效的化妆品原料组合物,其特征在于,包括以下成分:光果甘草根提取物、姜黄根提取物、黄芩根提取物、芍药根提取物、三七根提取物;所述光果甘草根提取物占组合物总浓度配比为5%,姜黄根提取物占组合物总浓度配比为5%,黄芩根提取物占组合物总浓度配比为26%,芍药根提取物占组合物总浓度配为11%,三七根提取物占组合物总浓度配比为53%。
2.根据权利要求1所述的具有美白舒缓功效的化妆品原料组合物,其特征在于:所述光果甘草根提取物对蘑菇酪氨酸酶活性的IC50为0.1μg/mL,姜黄根提取物对蘑菇酪氨酸酶活性的IC50为2μg/mL。
3.根据权利要求1所述的具有美白舒缓功效的化妆品原料组合物,其特征在于:黄芩根提取物、芍药根提取物和三七根提取物均可抑制NO的分泌,IC50分别为27μg/mL、41.6μg/mL、190μg/mL。
4.根据权利要求1所述的具有美白舒缓功效的化妆品原料组合物,其特征在于:所述光果甘草根和姜黄根提取物由体外蘑菇酪氨酸酶活性抑制试验筛选出;所述黄芩根提取物、芍药根提取物以及三七根提取物由LPS诱导巨噬细胞RAW264.7检测NO含量试验筛选出。
5.一种如权利要求1所述的具有美白舒缓功效的化妆品原料组合物的评价体系,其特征在于,包括:
高通量初筛功效模块,用于对所收集的美白、舒缓原料进行初步筛选;
基因、细胞功效评价模块,用于美白、舒缓原料组合物复配比例优化及功效评价;
3D皮肤模型功效评价模块,用于对含该原料组合物的化妆品成品进行功效评价。
6.根据权利要求5所述的评价体系,其特征在于:所述高通量初筛功效模块,采用体外蘑菇酪氨酸酶活性抑制试验筛选美白原料;采用LPS诱导巨噬细胞RAW264.7检测NO含量试验筛选舒缓原料。
7.根据权利要求5所述的评价体系,其特征在于:基因、细胞功效评价模块,包括对筛选出来的原料通过响应面法进行复配比例优化。
8.根据权利要求5所述的评价体系,其特征在于:所述美白、舒缓原料组合物功效评价具体为:基于α-MSH诱导小鼠黑素瘤细胞B16检测黑色素含量、酪氨酸酶基因表达量试验评价美白功效;基于UVB辐照角质形成细胞HaCat检测ROS含量试验,基于H2O2刺激角质形成细胞HaCat检测SOD、GPx活性和MDA含量试验评价抗氧化功效;基于LPS诱导巨噬细胞RAW264.7检测IL-6、TNF-α、NO含量试验评价舒缓功效。
9.根据权利要求5所述的评价体系,其特征在于:所述3D皮肤模型功效评价模块具体为:基于UVB刺激3D黑素模型检测表观色度、表观亮度试验评价美白功效,基于SLS刺激3D表皮皮肤模型检测IL-1α含量试验评价舒缓功效。
10.根据权利要求5所述的评价体系,其特征在于:还包括人体临床功效评价模块;所述人体临床功效评价模块具体为:基于紫外线诱导人体皮肤黑化模型检测黑素指数、ITA°及视觉肤色评估来进行美白临床功效评价。
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